Name: Anti-Caspase-3 antibody [EPR18297]
SKU: ab184787
Rating: 3 (2 reviews)

Anti-Caspase-3 antibody [EPR18297] is a rabbit recombinant monoclonal antibody that is used to detect Caspase-3 in IHC-P, IP, Western blot. Suitable for Human, Mouse, Rat samples.

- Using biophysical QC, antibody identity is confirmed at a molecular level for unrivaled batch-batch consistency
- Specificity confirmed with CASP3 knockout cell line validation
- Specificity and sensitivity confirmed in IHC with multi-tissue microarray (TMA) validation
- Antibody clone EPR18297 is cited in over 330 publications

Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 antibody [EPR18297] (AB184787), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	WB	IP	IHC-P
Human	Tested	Tested	Tested
Mouse	Tested	Expected	Expected
Rat	Tested	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/2000	Notes The 17 kDa band is the active form of the cleaved caspase 3 (subunit p17). ab184787 recognizes pro-Caspase 3 and unable to detect the active caspases after induction in mouse and rat samples. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Species Rat	Dilution info 1/2000	Notes The 17 kDa band is the active form of the cleaved caspase 3 (subunit p17). ab184787 recognizes pro-Caspase 3 and unable to detect the active caspases after induction in mouse and rat samples.
Species Human	Dilution info 1/2000	Notes The 17 kDa band is the active form of the cleaved caspase 3 (subunit p17). ab184787 recognizes pro-Caspase 3 and unable to detect the active caspases after induction in mouse and rat samples.

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/80	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/1000	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

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Target data

See full target information CASP3

Function

Thiol protease that acts as a major effector caspase involved in the execution phase of apoptosis (PubMed:18723680, PubMed:20566630, PubMed:23650375, PubMed:35338844, PubMed:35446120, PubMed:7596430). Following cleavage and activation by initiator caspases (CASP8, CASP9 and/or CASP10), mediates execution of apoptosis by catalyzing cleavage of many proteins (PubMed:18723680, PubMed:20566630, PubMed:23650375, PubMed:7596430). At the onset of apoptosis, it proteolytically cleaves poly(ADP-ribose) polymerase PARP1 at a '216-Asp-|-Gly-217' bond (PubMed:10497198, PubMed:16374543, PubMed:7596430, PubMed:7774019). Cleaves and activates sterol regulatory element binding proteins (SREBPs) between the basic helix-loop-helix leucine zipper domain and the membrane attachment domain (By similarity). Cleaves and activates caspase-6, -7 and -9 (CASP6, CASP7 and CASP9, respectively) (PubMed:7596430). Cleaves and inactivates interleukin-18 (IL18) (PubMed:37993714, PubMed:9334240). Involved in the cleavage of huntingtin (PubMed:8696339). Triggers cell adhesion in sympathetic neurons through RET cleavage (PubMed:21357690). Cleaves and inhibits serine/threonine-protein kinase AKT1 in response to oxidative stress (PubMed:23152800). Acts as an inhibitor of type I interferon production during virus-induced apoptosis by mediating cleavage of antiviral proteins CGAS, IRF3 and MAVS, thereby preventing cytokine overproduction (PubMed:30878284). Also involved in pyroptosis by mediating cleavage and activation of gasdermin-E (GSDME) (PubMed:35338844, PubMed:35446120). Cleaves XRCC4 and phospholipid scramblase proteins XKR4, XKR8 and XKR9, leading to promote phosphatidylserine exposure on apoptotic cell surface (PubMed:23845944, PubMed:33725486). Cleaves BIRC6 following inhibition of BIRC6-caspase binding by DIABLO/SMAC (PubMed:36758104, PubMed:36758106).

Alternative names

CPP32, CASP3, Caspase-3, CASP-3, Apopain, Cysteine protease CPP32, Protein Yama, SREBP cleavage activity 1, CPP-32, SCA-1

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EPR18297
Purification technique: Affinity purification Protein A
Specificity: This antibody recognizes pro-Caspase 3 and potentially cross reacts with active caspases after apoptosis has been induced in wildtype cells and not Caspase 3 knockout cells
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

Anti-Caspase-3 antibody [EPR18297] (ab184787) was developed by Abcam using patented rabbit monoclonal antibody technology and is validated for use in IHC-P, IP, WB in human, mouse, rat samples.

Anti-Caspase-3 antibody [EPR18297] (ab184787) has been cited over 223 times in peer reviewed journals and is trusted by the scientific community.

Abcam's high quality manufacturing and validation processes ensure Anti-Caspase-3 antibody [EPR18297] (ab184787) has high sensitivity and specificity alongside high lot-to-lot consistency and reproducibility.

The specificity of Anti-Caspase-3 antibody [EPR18297] (ab184787) has been confirmed by testing in knockout samples.

Anti-Caspase-3 antibody [EPR18297] (ab184787) specifically detects Caspase-3 (UniProt ID: P42574; Molecular weight: 17kDa) and is sold in a convenient trial size to enable initial testing (20 µL) and larger sizes for subsequent scaling up experiments (100 µL and 1 mL).

Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

This product is a recombinant monoclonal antibody, which offers several advantages including:

- High batch-to-batch consistency and reproducibility
- Improved sensitivity and specificity
- Long-term security of supply
- Animal-free batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
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10 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 antibody [EPR18297] (ab184787)
Immunohistochemical analysis of paraffin-embedded Human cervical cancer tissue labeling active and pro Caspase 3 with ab184787 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/500 dilution. Nucleus and cytoplasm staining on tumor cells of Human cervix cancer is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
Lane 1: Wild-type HAP1 cell lysate + Staurosporine (1μM for 4h)
Lane 2: Wild-type HAP1 cell lysate
Lane 3: Caspase-3 knockout HAP1 cell lysate + Staurosporine (1μM for 4h)
Lane 4: Caspase-3 knockout HAP1 cell lysate
Lanes 1 - 4: Merged signal (red and green). Green - ab184787 observed at 35 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.

ab184787 was shown to recognise pro Caspase 3 when Caspase 3 knockout samples were used, along with additional cross-reactive bands. Wild-type and Caspase 3 knockout samples (± Staurosporine treatment) were subjected to SDS-PAGE. ab184787 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (loading control to GAPDH) were diluted to 1/2000 and 1/10000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776)
secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
Predicted band size: 31 kDa
Observed band size: 32 kDa
Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
Blocking and diluting buffer: 5% NFDM/TBST.

ab184787 recognizes pro-Caspase 3 and unable to detect the active caspases after induction in mouse and rat samples.
All lanes: Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787) at 0.7 µg/mL
Lane 1: Untreated NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg
Lane 2: NIH/3T3 (Mouse embryonic fibroblast) treated with 1μM Staurosporine for 4 hours whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 31 kDa
Observed band size: 32 kDa
Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
Blocking/Dilution buffer: 5% NFDM/TBST.
Specificity: interacts with full length pro-Caspase 3 and the p17 subunit.
The Caspase-3 precursor is first cleaved between D175 and S176 to produce the p11 subunit and p20 fragment. Subsequently, the p20 fragment is cleaved between D28 and S29 to generate the p17 subunit (Proc. Natl. Acad. Sci. USA. 93, 7464-7469 - PMID:8755496).
All lanes: Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787) at 1/2000 dilution
Lane 1: Untreated Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates at 10 µg
Lane 2: Jurkat whole cell lysates treated with 1uM staurosporine for 4 hours at 10 µg
Secondary
All lanes: Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution
Predicted band size: 31 kDa
Observed band size: 17 kDa, 32 kDa
Exposure time: 1min
Immunoprecipitation - Anti-Caspase-3 antibody [EPR18297] (ab184787)
active and pro Caspase 3 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate treated with 1uM staurosporine for 4 hours with ab184787 at 1/80 dilution. Western blot was performed from the immunoprecipitate using ab184787 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1/1500 dilution.
Lane 1: HeLa whole cell lysate treated with 1uM staurosporine for 4 hours 10 μg (Input).
Lane 2: ab184787 IP in HeLa whole cell lysate treated with 1uM staurosporine for 4 hours.
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab184787 in HeLa whole cell lysate treated with 1uM staurosporine for 4 hours.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes.
All lanes: Immunoprecipitation - Anti-Caspase-3 antibody [EPR18297] (ab184787)
Predicted band size: 31 kDa
Observed band size: 17 kDa, 32 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 antibody [EPR18297] (ab184787)
Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling active and pro Caspase 3 with ab184787 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) secondary antibody at 1/500 dilution. Nucleus and cytoplasm staining on lymphocytes of tonsil is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as GAPDH loading control.

Bands between 27-32kDa represent cleavage of the procaspase at D9 and D28, respectively (PMID: 14567691)
All lanes: Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787) at 1/1000 dilution
Lane 1: Rat brain tissue lysate at 20 µg
Lane 2: Rat hippocampus tissue lysate at 20 µg
Lane 3: Rat spinal cord tissue lysate at 20 µg
Lane 4: Rat cerebellum tissue lysate at 20 µg
Lane 5: Rat cerebral cortex tissue lysate at 20 µg
Lane 6: Rat hypothalamus tissue lysate at 20 µg
Lane 7: Rat heart tissue lysate at 20 µg
Lane 8: Rat liver tissue lysate at 20 µg
Lane 9: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 31 kDa
Exposure time: 20s
Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
Anti-CASP3 antibody [EPR18297] (ab184787) staining at 1/2000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab184787 was shown to bind specifically to CASP3. A band was observed at 35 kDa in treated wild-type HAP1 and HeLa cell lysates with no signal observed at this size in CASP3 knockout HAP1 cell line and a band at a lower molecular weight in the CAPS3 knockout HeLa cell line Human CASP3 (Caspase-3) knockout HeLa cell line ab255370 (knockout cell lysate ab263779) which cannot be cleaved to active CASP3. To generate this image, wild-type and CASP3 knockout HAP1 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787) at 1/2000 dilution
Lane 1: Wild-type HAP1 Treated Staurosporine (2 uM, 4h) cell lysate at 20 µg
Lane 2: CASP3 knockout HAP1 Treated Staurosporine (2 uM, 4h) cell lysate at 20 µg
Lane 3: Wild-type HAP1 Vehicle Control Staurosporine (0 uM, 4h) cell lysate at 20 µg
Lane 4: CASP3 knockout HAP1 Vehicle Control Staurosporine (0 uM, 4h) cell lysate at 20 µg
Lane 5: Wild-type HeLa Treated Staurosporine (2 uM, 4h) cell lysate at 20 µg
Lane 6: CASP3 knockout HeLa Treated Staurosporine (2 uM, 4h) cell lysate at 20 µg
Lane 7: Wild-type HeLa Vehicle Control Staurosporine (0 uM, 4h) cell lysate at 20 µg
Lane 8: CASP3 knockout HeLa Vehicle Control Staurosporine (0 uM, 4h) cell lysate at 20 µg
Secondary
All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 31 kDa
Observed band size: 35 kDa
Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787)
Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as GAPDH loading control.

Bands between 27-32kDa represent cleavage of the procaspase at D9 and D28, respectively (PMID: 14567691)
All lanes: Western blot - Anti-Caspase-3 antibody [EPR18297] (ab184787) at 1/1000 dilution
Lane 1: Mouse Alzheimer's disease brain tissue lysate at 20 µg
Lane 2: Mouse brain cancer tissue lysate at 20 µg
Lane 3: Mouse hippocampus tissue lysate at 20 µg
Lane 4: Mouse spinal cord tissue lysate at 20 µg
Lane 5: Mouse cerebellum tissue lysate at 20 µg
Lane 6: Mouse cerebral cortex tissue lysate at 20 µg
Lane 7: Mouse hypothalamus tissue lysate at 20 µg
Lane 8: Mouse heart tissue lysate at 20 µg
Lane 9: Mouse liver tissue lysate at 20 µg
Lane 10: Human brain tissue lysate at 20 µg
Lane 11: Human liver tissue lysate at 20 µg
Lane 12: Human hypothalamus tissue lysate at 20 µg
Lane 13: Human heart tissue lysate at 20 µg
Lane 14: Human cerebellum tissue lysate at 20 µg
Secondary
All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 32 kDa
Exposure time: 10s
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Caspase-3 antibody [EPR18297] (ab184787)
Caspase-3 Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) staining of Human tonsil tissue using rabbit Anti-Caspase-3 antibody
Immunohistochemical analysis of formalin fixed paraffin embedded human tonsil labelling Caspase-3 with ab184787 at a concentration of 0.05µg/ml. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with a OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was performed with DISCOVERY cell conditioning solution (CC1) 100°C, pH8.5 for 32mins.
ab184787 Anti-Caspase-3 antibody [EPR18297] antibody was incubated for 16mins at 37°C. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
Customers are encouraged to optimise antigen retrieval conditions, antibody concentration, incubation times and temperature for best results in their own IHC assay workflow (automated and manual)